PUBLICATION

Molecular multi-effect screening of environmental pollutants using the MolDarT

Authors
Liedtke, A., Muncke, J., Rüfenacht, K., and Eggen, R.I.
ID
ZDB-PUB-080218-27
Date
2008
Source
Environmental toxicology   23(1): 59-67 (Journal)
Registered Authors
Keywords
developing zebrafish, immune disruption, detoxification, metal toxicity, molecular effect, rag1, metallothionein 2, cyp1a1
MeSH Terms
  • Animals
  • Cadmium Chloride/toxicity
  • Cytochrome P-450 CYP1A1/drug effects
  • Cytochrome P-450 CYP1A1/genetics
  • DNA Primers
  • Environmental Exposure*
  • Environmental Monitoring/methods
  • Environmental Pollutants/toxicity*
  • Estrogens/toxicity*
  • Gene Expression/drug effects
  • Naphthalenes/toxicity
  • RNA, Messenger/analysis
  • Reverse Transcriptase Polymerase Chain Reaction
  • Zebrafish/genetics*
  • Zebrafish Proteins/drug effects*
  • Zebrafish Proteins/genetics
  • Zinc Sulfate/toxicity
PubMed
18214934 Full text @ Env. Tox.
CTD
18214934
Abstract
Molecular effect detection is a useful approach for ecotoxicological screening of chemicals. We show here the application of the molecular DarT (MolDarT), where the expression of selected target genes is detected in short-term (120 h) exposed developing zebrafish (Danio rerio), thus allowing subacute multi-effect compound screening. The genes metallothionein 2 (mt2), cytochrome P450 1A1 (cyp1a1), and recombination activation gene 1 (rag1) are used as endpoints that describe detoxification/metal toxicity (mt2), detoxification/PAH toxicity (cyp1a1), and acquired immune system disruption (rag1). Each gene's developmental expression was studied in unexposed zebrafish during 4 to 120 h past fertilization (hpf), and all three genes were found to be expressed at 120 hpf. Furthermore, mt2 transcripts were present at high levels at 4 hpf, indicating a maternal transfer. For positive toxicity controls, freshly fertilized zebrafish eggs were exposed for 120 hpf to ZnSO(4), 1,5-dimethylnaphthalene (DMN) and CdCl(2). Exposure to 100 and 200 muM ZnSO(4) significantly induced mt2; 10 muM DMN and 20 muM DMN resulted in significantly increased cyp1a1 abundance; and 5 and 10 muM CdCl(2) significantly reduced rag1 expression levels. Furthermore, we analysed these target genes for their expression in zebrafish eggs from a previous exposure study. The eggs were exposed for 120 hpf to the environmental pollutants estradiol (E2), ethinylestradiol (EE2), nonylphenol (NP), atrazine, cyproconazol, and bisphenol A (BPA) and found differential expression of the three genes. Exposure to the (xeno-)estrogenic compound NP (0.75 muM) significantly lowered mt2 expression. This study shows the potential of short-term in vivo multi-effect screenings within one single subacute exposure using the MolDarT.
Genes / Markers
Figures
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Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping